A novel ACE2-Based electrochemical biosensor for sensitive detection of SARS-CoV-2.

SARS-CoV-2 emerged in late 2019 and quickly spread globally, resulting in significant morbidity, mortality, and socio-economic disruptions. As of now, collaborative global efforts in vaccination and the advent of novel diagnostic tools have considerably curbed the spread and impact of the virus in many regions. Despite this progress, the demand remains for low-cost, accurate, rapid and scalable diagnostic tools to reduce the influence of SARS-CoV-2. Herein, the angiotensin-converting enzyme 2 (ACE2), a receptor for SARS-CoV-2, was immobilized on two types of electrodes, a screen-printed gold electrode (SPGE) and a screen-printed carbon electrode (SPCE), to develop electrochemical biosensors for detecting SARS-CoV-2 with high sensitivity and selectivity. This was achieved by using 1H, 1H, 2H, 2H-perfluorodecanethiol (PFDT) and aryl diazonium salt serving as linkers for SPGEs and SPCEs, respectively. Once SARS-CoV-2 was anchored onto the ACE2, the interaction of the virus with the redox probe was analyzed using electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). Aryl diazonium salt was observed as a superior linker compared to PFDT due to its consistent performance in the modification of the SPCEs and effective ACE2 enzyme immobilization. A distinct pair of redox peaks in the cyclic voltammogram of the biosensor modified with aryl diazonium salt highlighted the redox reaction between the functional groups of SARS-CoV-2 and the redox probe. The sensor presented a linear relationship between the redox response and the logarithm of SARS-CoV-2 concentration, with a detection limit of 1.02 × 106 TCID50/mL (50% tissue culture infectious dose). Furthermore, the biosensor showed remarkable selectivity towards SARS-CoV-2 over H1N1virus.

Reduced Glutathione-Modified Electrode for the Detection of Hydroxyl Free Radicals.

Hydroxyl radicals (•OH) are known as essential chemicals for cells to maintain their normal functions and defensive responses. However, a high concentration of •OH may cause oxidative stress-related diseases, such as cancer, inflammation, and cardiovascular disorders. Therefore, •OH can be used as a biomarker to detect the onset of these disorders at an early stage. Reduced glutathione (GSH), a well-known tripeptide for its antioxidant capacity against reactive oxygen species (ROS), was immobilized on a screen-printed carbon electrode (SPCE) to develop a real-time detection sensor with a high selectivity towards •OH. The signals produced by the interaction of the GSH-modified sensor and •OH were characterized using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The CV curve of the GSH-modified sensor in the Fenton reagent exhibited a pair of well-defined peaks, demonstrating the redox reaction of the electrochemical sensor and •OH. The sensor showed a linear relationship between the redox response and the concentration of •OH with a limit of detection (LOD) of 49 µM. Furthermore, using EIS studies, the proposed sensor demonstrated the capability of differentiating •OH from hydrogen peroxide (H2O2), a similar oxidizing chemical. After being immersed in the Fenton solution for 1 hr, redox peaks in the CV curve of the GSH-modified electrode disappeared, revealing that the immobilized GSH on the electrode was oxidized and turned to glutathione disulfide (GSSG). However, it was demonstrated that the oxidized GSH surface could be reversed back to the reduced state by reacting with a solution of glutathione reductase (GR) and nicotinamide adenine dinucleotide phosphate (NADPH), and possibly reused for •OH detection.